The minimal virus requirements for the generation of influenza virus-like particle (VLP) assembly and budding were reassessed. Using neuraminidase (NA) from the H5N1 and H1N1 subtypes, it was found that the expression of NA alone was sufficient to generate and release VLPs. Biochemical and functional characterization of the NA-containing VLPs demonstrated that they were morphologically similar to influenza virions. The NA oligomerization was comparable to that of the live virus, and the enzymic activity, whilst not required for the release of NA-VLPs, was preserved. Together, these findings indicate that NA plays a key role in virus budding and morphogenesis, and demonstrate that NA-VLPs represent a useful tool in influenza research.


Article metrics loading...

Loading full text...

Full text loading...



  1. Air, G. M. & Laver, W. G.(1989). The neuraminidase of influenza virus. Proteins 6, 341–356.[CrossRef] [Google Scholar]
  2. Barman, S., Adhikary, L., Chakraborti, A. K., Bernas, C., Kawaoka, Y. & Nayak, D. P.(2004). Role of transmembrane domain and cytoplasmic tail amino acid sequences of influenza A virus neuraminidase in raft-association and virus budding. J Virol 78, 5258–5269.[CrossRef] [Google Scholar]
  3. Bucher, D. & Palese, P.(1975). The biologically active proteins of influenza virus: neuraminidase. In The Influenza Viruses and Influenza, pp. 83–123. Edited by E. Kilbourne. New York: Academic Press.
  4. Chen, B. J., Leser, G. P., Morita, E. & Lamb, R. A.(2007). Influenza virus hemagglutinin and neuraminidase, but not the matrix protein, are required for assembly and budding of plasmid-derived virus-like particles. J Virol 81, 7111–7123.[CrossRef] [Google Scholar]
  5. França de Barros, J., Jr, Sales Alviano, D., da Silva, M. H., Dutra Wigg, M., Sales Alviano, C., Schauer, R. & dos Santos Silva Couceiro, J. N.(2003). Characterization of sialidase from an influenza A (H3N2) virus strain: kinetic parameters and substrate specificity. Intervirology 46, 199–206.[CrossRef] [Google Scholar]
  6. Fujiyoshi, Y., Kume, N. P., Sakata, K. & Sato, S. B.(1994). Fine structure of influenza A virus observed by electron cryo-microscopy. EMBO J 13, 318–326. [Google Scholar]
  7. Gómez-Puertas, P., Albo, C., Pérez-Pastrana, E., Vivo, A. & Portela, A.(2000). Influenza virus matrix protein is the major driving force in virus budding. J Virol 74, 11538–11547.[CrossRef] [Google Scholar]
  8. Huang, I. C., Li, W., Sui, J., Marasco, W., Choe, H. & Farzan, M.(2008). Influenza A virus neuraminidase limits viral superinfection. J Virol 82, 4834–4843.[CrossRef] [Google Scholar]
  9. Jin, H., Leser, G. P., Zhang, J. & Lamb, R. A.(1997). Influenza virus hemagglutinin and neuraminidase cytoplasmic tails control particle shape. EMBO J 16, 1236–1247.[CrossRef] [Google Scholar]
  10. Latham, T. & Galaeza, J. M.(2001). Formation of wild-type and chimeric influenza virus-like particles following simultaneous expression of only four structural proteins. J Virol 75, 6154–6165.[CrossRef] [Google Scholar]
  11. Liu, C., Eichelberger, M. C., Compans, R. W. & Air, G. M.(1995). Influenza type A virus neuraminidase does not play a role in viral entry, replication, assembly, or budding. J Virol 69, 1099–1106. [Google Scholar]
  12. Matrosovich, M. N., Matrosovich, T. Y., Gray, T., Roberts, N. A. & Klenk, H. D.(2004). Neuraminidase is important for the initiation of influenza virus infection in human airway epithelium. J Virol 78, 12665–12667.[CrossRef] [Google Scholar]
  13. Matrosovich, M. N., Klenk, H. D. & Kawaoka, Y.(2006). Receptor specificity, host range and pathogenicity of influenza viruses. In Influenza Virology: Current Topics, pp. 95–137. Edited by Y. Kawaoka. Wymondham, UK: Caister Academic Press.
  14. Mochalova, L., Kurova, V., Shtyrya, Y., Korchagina, E., Gambaryan, A., Belyanchikov, I. & Bovin, N.(2007). Oligosaccharide specificity of influenza H1N1 virus neuraminidases. Arch Virol 152, 2047–2057.[CrossRef] [Google Scholar]
  15. Nagai, T., Suzuki, Y. & Yamada, H.(1995). Comparison of substrate specificities of sialidase activity between purified enzymes from influenza virus A (H1N1 and H3N2 subtypes) and B strains and their original viruses. Biol Pharm Bull 18, 1251–1254.[CrossRef] [Google Scholar]
  16. Nayak, D. P., Balogun, R. A., Yamada, H., Zhou, Z. H. & Barman, S.(2009). Influenza virus morphogenesis and budding. Virus Res 143, 147–161.[CrossRef] [Google Scholar]
  17. Neumann, G., Watanabe, T. & Kawaoka, Y.(2000). Plasmid-driven formation of influenza virus-like particles. J Virol 74, 547–551.[CrossRef] [Google Scholar]
  18. Nicholls, J. M., Bourne, A. J., Chen, H., Guan, Y. & Peiris, J. S.(2007). Sialic acid receptor detection in the human respiratory tract: evidence for widespread distribution of potential binding sites for human and avian influenza viruses. Respir Res 8, 73[CrossRef] [Google Scholar]
  19. Noda, T., Sagara, H., Yen, A., Takada, A., Kida, K., Cheng, R. H. & Kawaoka, Y.(2006). Architecture of ribonucleoprotein complexes in influenza A virus particles. Nature 439, 490–492.[CrossRef] [Google Scholar]
  20. Ohuchi, M., Asaoka, N., Sakai, T. & Ohuchi, R.(2006). Roles of neuraminidase in the initial stage of influenza virus infection. Microbes Infect 8, 1287–1293.[CrossRef] [Google Scholar]
  21. Saito, T., Taylor, G. & Webster, R. G.(1995). Steps in maturation of influenza A virus neuraminidase. J Virol 69, 5011–5017. [Google Scholar]
  22. Su, B., Wurtzer, S., Rameix-Welti, M. A., Dwyer, D., van der Werf, S., Naffakh, N., Clavel, F. & Labrosse, B.(2009). Enhancement of the influenza A hemagglutinin (HA)-mediated cell–cell fusion and virus entry by the viral neuraminidase (NA). PLoS One 4, e8495[CrossRef] [Google Scholar]
  23. Tscherne, D. M., Manicassamy, B. & Garcia-Sastre, A.(2010). An enzymatic virus-like particle assay for sensitive detection of virus entry. J Virol Methods 163, 336–343.[CrossRef] [Google Scholar]
  24. Wang, D., Harmon, A., Jin, J., Francis, D. H., Christopher-Hennings, J., Nelson, E., Montelaro, R. C. & Li, F.(2010). The lack of an inherent membrane targeting signal is responsible for the failure of the matrix (M1) protein of influenza A virus to bud into virus-like particles. J Virol 84, 4673–4681.[CrossRef] [Google Scholar]
  25. Zhang, J., Pekosz, A. & Lamb, R. A.(2000). Influenza virus assembly and lipid raft microdomains: a role for the cytoplasmic tails of the spike glycoproteins. J Virol 74, 4634–4644.[CrossRef] [Google Scholar]

Data & Media loading...


vol. , part 9, pp. 2322 - 2330

TEM visualization of influenza NA VLP formation

Desialylation of cell-surface sialylglycan by NA-VLPs

Comparative expression and release of NA-VLP for different NAs [Single PDF file](641 KB)


Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error